Objective We report a case of application of third-generation sequencing(TGS)combined with preimplantation genetic testing(PGT)for successful prevention of hereditary spastic paraplegia(HSP)caused by SPAST gene mutations and assess the value of PGT-M and TGS in managing hereditary spastic paraplegia.Methods A family affected by HSP underwent whole exon sequencing(WES),and a c.1699G>T mutation in the SPAST gene was identified.The mutation site in the proband was confirmed through Sanger sequencing.A single nucleotide polymorphism(SNP)site flanking the SPAST gene mutation was selected as the genetic linkage marker,and a SNP haplotype carrying the mutated gene was constructed.Ovarian stimulation using an antagonist regimen was performed for oocyte retrieval,followed by intracytoplasmic sperm injection(ICSI)and embryo culture.Blastocyst trophectoderm cells were biopsied for preimplantation genetic testing for monogenic disorders(PGT-M)to allow the selection of disease-free embryos for transfer.Results In this cycle,a total of 20 oocytes were retrieved,among which 18 were successfully fertilized to result in 12 blastocysts eligible for biopsy.Genetic testing revealed that all the 12 blastocysts were successfully amplified and confirmed as euploidy.Among them,8 blastocysts did not carry paternal mutations,and a high-quality euploid embryo was selected for frozen embryo transfer(FET).Subsequent amniotic fluid testing during pregnancy confirmed the absence of paternal mutations in the fetus,resulting in the birth of a healthy baby girl.Conclusion For cases of genetic diseases with missing pedigree data,the application of third-generation sequencing and PGT-M technique can effectively block vertical transmission of SPAST gene mutation to the offspring,avoid pregnancy with an aneuploid embryo,and help families with autosomal dominant HSP obtain healthy offsprings.

With the increasing life expectancy and lifestyle changes of patients with chronic hepatitis B (CHB), the significance of comorbidities of chronic non-communicable diseases (NCDs) in disease progression and health prognosis of CHB patients is gaining prominence. This study aims to explore the association between CHB and NCDs comorbidities, focusing on the impact of common metabolism-related diseases, such as metabolic syndrome and diabetes, on the health outcomes of CHB patients. We also summarize studies on integrating the management of comorbidities in CHB patients and provide relevant recommendations for effective management. The findings of this study serve as a foundation for understanding the clinical characteristics and prevalence trends, reducing the disease burden of comorbidities among CHB patients, and establishing a comprehensive and coordinated management system for comorbidities.

Objective We report a case of application of third-generation sequencing(TGS)combined with preimplantation genetic testing(PGT)for successful prevention of hereditary spastic paraplegia(HSP)caused by SPAST gene mutations and assess the value of PGT-M and TGS in managing hereditary spastic paraplegia.Methods A family affected by HSP underwent whole exon sequencing(WES),and a c.1699G>T mutation in the SPAST gene was identified.The mutation site in the proband was confirmed through Sanger sequencing.A single nucleotide polymorphism(SNP)site flanking the SPAST gene mutation was selected as the genetic linkage marker,and a SNP haplotype carrying the mutated gene was constructed.Ovarian stimulation using an antagonist regimen was performed for oocyte retrieval,followed by intracytoplasmic sperm injection(ICSI)and embryo culture.Blastocyst trophectoderm cells were biopsied for preimplantation genetic testing for monogenic disorders(PGT-M)to allow the selection of disease-free embryos for transfer.Results In this cycle,a total of 20 oocytes were retrieved,among which 18 were successfully fertilized to result in 12 blastocysts eligible for biopsy.Genetic testing revealed that all the 12 blastocysts were successfully amplified and confirmed as euploidy.Among them,8 blastocysts did not carry paternal mutations,and a high-quality euploid embryo was selected for frozen embryo transfer(FET).Subsequent amniotic fluid testing during pregnancy confirmed the absence of paternal mutations in the fetus,resulting in the birth of a healthy baby girl.Conclusion For cases of genetic diseases with missing pedigree data,the application of third-generation sequencing and PGT-M technique can effectively block vertical transmission of SPAST gene mutation to the offspring,avoid pregnancy with an aneuploid embryo,and help families with autosomal dominant HSP obtain healthy offsprings.

Objective : To investigate the expression characteristics of mu opioid receptor ( MOR) in rat colon smooth muscle cells by cultured rat primary colonic smooth muscle cells . Methods : colonic smooth muscle cells were isolated , cultured and identified ; immunofluorescence double labeling method was used to observe the distribution characteristics of MOR , Endomorphin⁃2 (EM2) , and calmodulin (CaM) in colonic smooth muscle cells ; Western Blot method was used to detect the expression of MOR and EM2 in smooth muscle cells of the colon . After the intervention measures Acetylcholine (ACh) ( 1 × 10 - 3 mol/L) and EM2 (2 μmol/L) were applied , the changes of CaM protein expression were observed ; The calciumion imaging method was used to detect the changes of calciumi on concentration in smooth muscle cells. Results: The colonic smooth muscle cells were cultured and identified . The positive cells labeled with α ⁃smooth muscle actin ( α ⁃SMA) accounted for more than 95% of the total number of cells . Immunofluorescence double labeling showed that there were MOR and EM2 distributions in colonic smooth muscle cells , and all MOR and EM2 positive cells coexisted with α ⁃SMA . Western Blot results showed that there were MOR and EM2 expressions in colonic smooth muscle cells . CaM in the ACh group significantly increased at 10 minutes (P < 0. 05) , CaM in the EM2 group significantly decreased ( P < 0. 05) ; The calciumion imaging results showed that alone applied ACh , the calciumion concentration in smooth muscle cells significantly increased ( P <0. 05) ; Alone applied EM2 , the calciumion concentration in colonic smooth muscle cells was down⁃regulated (P <0. 05) ; Applied ACh and EM2 sequentially , EM2 significantly reduced the increase of the calciumion concentration in smooth muscle cells induced by ACh (P < 0. 05) . Conclusion MOR and EM2 are expressed in colonic smooth muscle cells , and EM2 may inhibit the expression of CaM and reduce the concentration of calcium ions through MOR .

Electrocardiogram (ECG) signal is an important basis for the diagnosis of arrhythmia and myocardial infarction. In order to further improve the classification effect of arrhythmia and myocardial infarction, an ECG classification algorithm based on Convolutional vision Transformer (CvT) and multimodal image fusion was proposed. Through Gramian summation angular field (GASF), Gramian difference angular field (GADF) and recurrence plot (RP), the one-dimensional ECG signal was converted into three different modes of two-dimensional images, and fused into a multimodal fusion image containing more features. The CvT-13 model could take into account local and global information when processing the fused image, thus effectively improving the classification performance. On the MIT-BIH arrhythmia dataset and the PTB myocardial infarction dataset, the algorithm achieved a combined accuracy of 99.9% for the classification of five arrhythmias and 99.8% for the classification of myocardial infarction. The experiments show that the high-precision computer-assisted intelligent classification method is superior and can effectively improve the diagnostic efficiency of arrhythmia as well as myocardial infarction and other cardiac diseases.
Humans ; Electrocardiography ; Heart Diseases ; Myocardial Infarction/diagnostic imaging* ; Algorithms ; Electric Power Supplies

Urea transporters (UT) play a vital role in the mechanism of urine concentration and are recognized as novel targets for the development of salt-sparing diuretics. Thus, UT inhibitors are promising for development as novel diuretics. In the present study, a novel UT inhibitor with a diarylamide scaffold was discovered by high-throughput screening. Optimization of the inhibitor led to the identification of a promising preclinical candidate,

Objective:To analyze the epidemiological characteristics of outbreaks of dengue fever in China from 2015 to 2018, and provide evidence for the prevention and control of dengue fever.Methods:We extracted the incidence data of dengue fever from China Disease Prevention and Control Information System, Public Health Emergency Reporting Management Information System and Vector Biological Monitoring System, and explored the epidemiological characteristics of the outbreaks in the past four years. Excel 2010 software and SPSS 20.0 software were used for data processing and analysis, ArcGIS 10.5 software was used for mapping.Results:A total of 111 outbreaks of dengue fever were reported nationwide from 2015 to 2018, involving 12 490 cases, accounting for 73.7% of the total cases in China. These outbreaks occurred in 85 counties and districts of 4 provinces, namely Guangdong (77 outbreaks), Yunnan (14 outbreaks), Zhejiang (8 outbreaks) and Fujian (8 outbreaks). The outbreaks occurred during May-November. Small-scale outbreaks with no more than 10 cases ended within 30 days (28/34, 82.4%) and larger-scale outbreaks lasted for several months. Dengue virus type 1 and type 2 were the main epidemic pathogens of dengue fever outbreaks in China. The outbreaks mainly occurred in areas with high population density and poor sanitary environment. There were significant differences in the age and occupational composition of the cases in the main outbreak provinces.Conclusions:Outbreaks of dengue fever can been seen in more areas in China, even in high latitudes areas. The epidemiologic characteristics of the outbreaks were different among provinces, showing as port type, rural type and urban type. Each province should adjust the control strategies accordingly.

Objective:To understand the epidemiological characteristics and risk factors of fatal cases of hand, foot, and mouth disease (HFMD) in children under 5 years old in China from 2008 to 2018, and provide evidence for the development of targeted prevention and control measures and reduction of the incidence of fatal HFMD cases.Methods:The incidence data of reported HFMD cases in China during 2008-2018 were collected from the National Notifiable Disease Surveillance Reporting System of China for the analyses on the demographic characteristics, spatial distribution, diagnosis or reporting and pathogen spectrum of the HFMD cases. Then the risk factors causing deaths were analyzed by using logistic regression model.Results:From 2008 to 2018, a total of 3 646 fatal cases of HFMD in children under 5 years old were reported in China. There were more fatal HFMD cases in boys than in girls (1.82∶1), the death mainly occurred in age group 0 to 2 years (87.71%). Adjusted mortality rate of HFMD in children under 5 years old in China declined from 0.87 per 100 000 in 2010 to 0.11 per 100 000 in 2018 (APC=-23.20%). In the 2 523 laboratory-confirmed deaths, 2 323 (92.07%) were EV-A71 infections, but the constituents of CV-A16 and other enterovirus infections increased. The interval from onset to diagnosis M=2( P25-P75:2 -4)d. The interval from onset to death M=3( P25-P75:2 -4)d. Age between 0 and 1 years, EV-A71 infection, longer interval between onset and diagnosis, and living in rural area were the risk factors causing fatal HFMD cases. Conclusions:The number of the fatal cases, the rate of mortality and case fatality HFMD in China had shown downward trends since 2010. EV-A71 is still the main pathogen causing fatal cases, but we should pay more attention to gene pattern of the other enteroviruses except EV-A71 and CV-A16. To reduce the risk of the fatal cases we should strengthen the health education about the immunization of EV-A71 inactivated vaccines and reduce the interval from onset to diagnosis in young children in western provinces and rural areas.

The oral microbiota is associated with oral diseases and digestive systemic diseases. Nevertheless, the causal relationship between them has not been completely elucidated, and colonisation of the gut by oral bacteria is not clear due to the limitations of existing research models. The aim of this study was to develop a human oral microbiota-associated (HOMA) mouse model and to investigate the ecological invasion into the gut. By transplanting human saliva into germ-free (GF) mice, a HOMA mouse model was first constructed. 16S rRNA gene sequencing was used to reveal the biogeography of oral bacteria along the cephalocaudal axis of the digestive tract. In the HOMA mice, 84.78% of the detected genus-level taxa were specific to the donor. Principal component analysis (PCA) revealed that the donor oral microbiota clustered with those of the HOMA mice and were distinct from those of specific pathogen-free (SPF) mice. In HOMA mice, OTU counts decreased from the stomach and small intestine to the distal gut. The distal gut was dominated by Streptococcus, Veillonella, Haemophilus, Fusobacterium, Trichococcus and Actinomyces. HOMA mice and human microbiota-associated (HMA) mice along with the GF mice were then cohoused. Microbial communities of cohoused mice clustered together and were significantly separated from those of HOMA mice and HMA mice. The Source Tracker analysis and network analysis revealed more significant ecological invasion from oral bacteria in the small intestines, compared to the distal gut, of cohoused mice. In conclusion, a HOMA mouse model was successfully established. By overcoming the physical and microbial barrier, oral bacteria colonised the gut and profiled the gut microbiota, especially in the small intestine.
Animals ; Bacteria ; Gastrointestinal Microbiome ; Germ-Free Life ; Humans ; Mice ; Microbiota ; RNA, Ribosomal, 16S

Objectives: To understand the status of studies about influenza economic burden in mainland China and summarize their major results. Methods: The words of influenza, flu, cost, economic, burden, effectiveness, benefit, utility, China, and Chinese, were used as search keywords. Journal papers published during 2000-2018 were searched from Chinese electronic databases (CNKI and Wanfang) and English electronic databases (PubMed, Web of science, EconLit and Cochrane Library). The language of literature was restricted to Chinese and English. A total of 23 effective documents were included, and the descriptive characteristics, research indexes and methods included in the literature were analyzed. The monetary unit used in this review is Chinese Yuan (CNY). Results: The 23 study sites were mainly in the relatively developed and populous regions. The total cost per capita of laboratory-confirmed influenza,of all age-group was reported in 6 literatures, and only 4 literatures reported it in out-patients (range: 768.0-999.9 CNY), Only one study reported this indicator in inpatients (9 832.0 CNY). One literature reported the total cost per capita of influenza-like illness,, which was 205.1 CNY. And one literature reported that the direct medical cost of inpatients per capita in children under 5 years of age was 6 072.0 CNY while two literature reported this index for the elderly over 60 years of age, ranging from 14 250.0 to 19 349.1 CNY. Four articles reported the economic burden of influenza in urban and rural areas, one of which showed that the related expenses of urban influenza inpatients accounted for 31% of the average annual income, while which for the rural flow was 113%. Conclusion The average economic burden of lab-confirmed influenza case is higher than that of influenza-like illness, and there are differences in outpatient indirect expenses and inpatients direct medical expenses. The direct medical burden for the hospitalized 60-years-and-beyond influenza case group is heavier thar other age group. By region, the influenza associated individual economic burden in rural area is higher than that of urban area..